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The number of nucleated cells (NC) and cell viability of pre-and post-SECCS bone marrow. (A,B) The number of NC in post-SECCS bone marrow was statistically lower than that in pre-SECCS bone marrow (Z = −2.482, P = 0.013). (C,D) There was no difference between the cell viability of pre-and post-SECCS bone marrow (t = 0.884, P = 0.382). NC (or cell viability) difference: NC number (or cell viability) difference between pre-and post-SECCS bone marrow samples.
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Bone defects are a common challenge in clinic, usually warranting bone grafts. However, current strategies to obtain effective graft materials have many drawbacks. Mesenchymal stem cell (MSC)-based therapy is a promising alternative. We designed an innovative appliance named the stem cell screen–enrich–combine(-biomaterials) circulating system (SEC...
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Context 1
... also found that the number of BMSCs varied greatly between different individuals, i.e., the BMSC pool in bone marrow was affected by multiple factors. As expected, MSC number tends to decrease with age (Fig. 3B). Besides, the proliferative capacity and osteoblast differentiation potential of BMSCs also reduce during aging 33 . Therefore, the curative effect of autologous MSC therapy may be compromised in elderly patients. Notably, patients with fresh fractures had more MSCs in the bone marrow reservoir (Fig. 3A). This can be explained by MSC ...
Context 2
... MSC number tends to decrease with age (Fig. 3B). Besides, the proliferative capacity and osteoblast differentiation potential of BMSCs also reduce during aging 33 . Therefore, the curative effect of autologous MSC therapy may be compromised in elderly patients. Notably, patients with fresh fractures had more MSCs in the bone marrow reservoir (Fig. 3A). This can be explained by MSC mobilization due to fresh fracture 34,35 . MSC mobilization promotes bone marrow cell proliferation, increasing the number of BMSC in the bone marrow. Meanwhile, MSC mobilization also enhances MSC homing (MSC are recruited from bone marrow niche and migrate to injured sites), increasing the number of ...
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Millions of people around the world have bone-tissue defects. Autologous and allogeneic bone grafting are frequent therapeutic techniques; however, none has produced the best therapeutic results. This has inspired researchers to investigate novel bone-regeneration technologies. In recent years, the development of bone tissue engineering (BTE) scaff...
A preclinical evaluation using a regenerative medicine methodology comprising an additively manufactured medical-grade ε-polycaprolactone β-tricalcium phosphate (mPCL-TCP) scaffold with a corticoperiosteal flap was undertaken in eight sheep with a tibial critical-size segmental bone defect (9.5 cm3, M size) using the regenerative matching axial vas...
Citations
... The Screen-Enrich-Combine(-Biomaterials) Circulating System also allows intraoperative enrichment of scaffold with autologous MSCs in 10-15 min. The device is based on the circulation of bone marrow through a porous β-TCP scaffold that acts as a filter to which MSCs adhere [74]. Wang et al. showed that besides MSCs platelets also adhered to porous β-TCP using Screen-Enrich-Combine Circulating System [75]. ...
Transfer of regenerative approaches into clinical practice is limited by strict legal regulation of in vitro expanded cells and risks associated with substantial manipulations. Isolation of cells for the enrichment of bone grafts directly in the Operating Room appears to be a promising solution for the translation of biomedical technologies into clinical practice. These intraoperative approaches could be generally characterized as a joint concept of tissue engineering in situ. Our review covers techniques of intraoperative cell isolation and seeding for the creation of tissue-engineered grafts in situ, that is, directly in the Operating Room. Up-to-date, the clinical use of tissue-engineered grafts created in vitro remains a highly inaccessible option. Fortunately, intraoperative tissue engineering in situ is already available for patients who need advanced treatment modalities.
... Mesenchymal stem cell (MSC)-based constructs have been considered promising alternatives to autologous bone grafts in repairing bone defects [1,2]. MSCs are multipotent stem cells and can differentiate into diverse cell types, such as osteoblasts, chondrocytes, and adipocytes under specific stimuli from culture media or biomaterials [3,4]. ...
Human umbilical cord mesenchymal stem cells (hUCMSCs) are promising for bone tissue engineering, which have a non-invasive harvesting process, high cell yield, favorable proliferation capacity, and low immunogenicity. However, the osteogenic efficacy of hUCMSCs is relatively lower than that of bone marrow mesenchymal stem cells (BMSCs). Hydrogels from decellularized extracellular matrix (dECM) preserve the biological compositions and functions of natural ECM, which can provide tissue-specific cues to regulate phenotypic expression and cell fate. It is unknown, however, whether hydrogels from periosteum can serve as pro-osteogenic carriers of hUCMSCs. Herein, a decellularized periosteum-derived hydrogel (dPH) was fabricated to reveal the effects of periosteum-specific cues on the bioactivities of hUCMSCs. A widely used non-bone/periosteum-derived ECM hydrogel product, Matrigel, was used as the control group. After decellularization, the absence of nuclei in the histological analysis indicated a successful removal of cellular components, which was also confirmed by DNA content quantification. The storage modulus of dPH increased (from 164.49 ± 29.92 Pa to 855.20 ± 20.67 Pa) with increasing concentration (from 0.5% to 1%). With a highly porous, fibrous microstructure, dPH had a more hydrophilic surface than Matrigel, of which the water contact angle reduced 62.62 ± 0.04%. Furthermore, dPH prominently promoted the initial cellular spreading with a significantly higher cell surface area (1.47-fold), cell spreading length (1.45-fold) and proliferation (approximately 1.05–1.13-fold) of hUCMSCs than those of Matrigel. Additionally, dPH was conducive to cell migration, whereas no cells migrated to Matrigel in the Transwell model. Compared with those of the Matrigel group, the osteogenesis-related genes expression levels (runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteopontin (OPN), and osteocalcin (OCN)) and mineralized matrix formation (9.74-fold) of the hUCMSCs significantly increased in the dPH group. Our study indicated that dPH could provide a pro-osteogenic microenvironment for hUCMSCs, thereby revealing a promising application potential to repair bone defects.
... The mean healing time for nonunion, fresh fracture, and other patients was 6.29, 3.12, and 4.72 months, respectively. The results showed that SECCS, which peri-operatively produces a bioactive composite of MSCs and β-TCP without in vitro culture, may represent a low cost and safe method for bone repair [151]. ...
Mesenchymal stem cell-based therapies are promising tools for bone tissue regeneration. However, tracking cells and maintaining them in the site of injury is difficult. A potential solution is to seed the cells onto a biocompatible scaffold. Construct development in bone tissue engineering is a complex step-by-step process with many variables to be optimized, such as stem cell source, osteogenic molecular factors, scaffold design, and an appropriate in vivo animal model. In this review, an MSC-based tissue engineering approach for bone repair is reported. Firstly, MSC role in bone formation and regeneration is detailed. Secondly, MSC-based bone tissue biomaterial design is analyzed from a research perspective. Finally, examples of animal preclinical and human clinical trials involving MSCs and scaffolds in bone repair are presented.
... At present, the insufficient amount of autologous bone and limited osteogenic ability of conventional bone substitutes has led to research efforts aimed at identifying bone fillings with excellent osteogenic properties (7)(8)(9). The stem cell Screen-Enrich-Combine(-biomaterials) Circulating System (SECCS), developed previously by us, can rapidly prepare enriched mesenchymal stem cell (MSCs)/beta tricalcium phosphate (β-TCP) composites, which have shown good bone repair ability in animal and various clinical bone defect models (10,11). However, the efficacy of repair needs to be further verified as an in-frame filling to repair complex anatomical bone defects. ...
Background:
Repairing complex anatomical load-bearing bone defects is difficult because it requires the restoration of the load-bearing function, reconstructing the anatomical shape, and repair by regenerated bone. We previously developed a Screen-Enrich-Combine(-biomaterials) Circulating System (SECCS) for rapid intraoperative enrichment of autologous bone marrow mesenchymal stem cells (MSCs) to enhance the osteogenic ability of porous bone substitutes. In this study, we prepared a 3D-printed Ti6A14V macroporous frame matching the defect shape to provide early load-bearing support and evaluated the efficacy of filling the frame with SECCS-processed MSCs/beta tricalcium phosphate (β-TCP) for long-term bone growth.
Methods:
Fifteen 2-year-old goats were involved in this study, and the lateral part of their distal femur was removed by an electric saw and was fitted by a matching electron beam melting technology-prepared (EBM) Ti6Al4V frame. Three types of frames, filled with nothing, pure porous β-TCP, or SECCS-processed MSCs/β-TCP, were fixed onto the defect site. Repair efficacy was evaluated by X-ray radiography, computed tomography (CT), histology, and histomorphometry.
Results:
In the basic regular hexagon printing unit, the combined side width (w) and the inscribed circle diameter (d) determines the printing frame's mechanical strength. The compressive load was significantly higher for w=1.9 mm, d=4.4 mm than for w=1.7 mm, d=4.0 mm or w=2.0 mm, d=5.0 mm (P<0.05). The EBM-prepared Ti6Al4V defect-matched frame was well maintained 9 months after implantation. The MSCs successfully adhered to the wall of the porous β-TCP in the SECCS-processed group and had spread fully in the test samples. Each goat in the MSCs/β-TCP-the filled group, had approximately 31,321.7±22,554.7 of MSCs and a larger area of new bone growth inside the frame than the control and blank areas groups.
Conclusions:
Filling the 3D-printed Ti6Al4V large-aperture frame with osteogenic materials achieved biological reconstruction over a larger area of regenerated bone to repair complex anatomical weight-bearing bone defects under the condition of early frame-supported load bearing. MSCs/β-TCP prepared by SECCS can be used as a filling material for this type of bone defect to obtain more efficacious bone repair.
... Bone marrow mesenchymal stem cells (BMSCs) have commonly been used in both basic studies [3,4] and clinical trials [5,6] as useful therapeutic seed cells for AMI. In addition to bone marrow, human placenta mesenchymal stem cells (PMSCs) transplantation were also considered to be an effective therapeutic approach for autoimmune diseases, bone repair, and cancer [7,8,9]. Human chorionic mesenchymal stem/stromal cells (CMSCs), also known as PMSCs, were isolated from third trimester placentas and were found to possess stem cell-like properties [10,11,12,13]. ...
Acute myocardial infarction (AMI) is the most critical heart disease. Mesenchymal stem cells (MSCs) have been widely used as a therapy for AMI for several years. The human placenta has emerged as a valuable source of transplantable cells of mesenchymal origin that can be used for multiple cytotherapeutic purposes. However, the different abilities of first trimester placental chorion mesenchymal stem cells (FCMSCs) and third trimester placental chorion mesenchymal stem cells (TCMSCs) have not yet been explored. In this study, we aimed to compare the effectiveness of FCMSCs and TCMSCs on the treatment of AMI. FCMSCs and TCMSCs were isolated and characterized, and then they were subjected to in vitro endothelial cell (EC) differentiation induction and tube formation to evaluate angiogenic ability. Moreover, the in vivo effects of FCMSCs and TCMSCs on cardiac improvement were also evaluated in a rat MI model. Both FCSMCs and TCMSCs expressed a series of MSCs surface markers. After differentiation induction, FCMSCs-derived EC (FCMSCs-EC) exhibited morphology that was more similar to that of ECs and had higher CD31 and vWF levels than TCMSCs-EC. Furthermore, tube formation could be achieved by FCMSCs-EC that was significantly better than that of TCMSCs-EC. Especially, FCMSCs-EC expressed higher levels of pro-angiogenesis genes, PDGFD, VEGFA, and TNC, and lower levels of anti-angiogenesis genes, SPRY1 and ANGPTL1. In addition, cardiac improvement, indicated by left ventricular end-diastolic diameter (LVEDd), left ventricular end-systolic diameter (LVEDs), left ventricular ejection fraction (LVEF) and left ventricular shortening fraction (LVSF), could be observed following treatment with FCMSCs, and it was superior to that of TCMSCs and Bone marrow MSCs (BMSCs). FCMSCs exhibited a superior ability to generate EC differentiation, as evidenced by in vitro morphology, angiogenic potential and in vivo cardiac function improvement; further, increased levels of expression of pro-angiogenesis genes may be the mechanism by which this effect occurred.
... Although these data are difficult to compare to the study of Hernigou, the application of the vacuum filter handle for 10 min during surgery could not harvest the same amount of progenitor cells as isolated by bone marrow aspiration [15]. However, the surgical vacuum suction device guarantes wetting of the bone substitute as well as cellular retaining without any additional invasive procedure and without further tissue or cell processing [17,37]. It is unclear whether a longer application of the vacuum is able to increase the amount of MNC in the filter biomaterial. ...
... Besides the application site (including cellular components) [37][38][39], the type of vacuum (level, duration) [17,37], the suction handle's material [38,39] and the type of bone substitute [40,41] might have an impact on in vitro results. ...
... Besides the application site (including cellular components) [37][38][39], the type of vacuum (level, duration) [17,37], the suction handle's material [38,39] and the type of bone substitute [40,41] might have an impact on in vitro results. ...
... Although these data are difficult to compare to the study of Hernigou, the application of the vacuum filter handle for 10 min during surgery could not harvest the same amount of progenitor cells as isolated by bone marrow aspiration [15]. However, the surgical vacuum suction device guarantes wetting of the bone substitute as well as cellular retaining without any additional invasive procedure and without further tissue or cell processing [17,37]. It is unclear whether a longer application of the vacuum is able to increase the amount of MNC in the filter biomaterial. ...
... Besides the application site (including cellular components) [37][38][39], the type of vacuum (level, duration) [17,37], the suction handle's material [38,39] and the type of bone substitute [40,41] might have an impact on in vitro results. ...
... Besides the application site (including cellular components) [37][38][39], the type of vacuum (level, duration) [17,37], the suction handle's material [38,39] and the type of bone substitute [40,41] might have an impact on in vitro results. ...
During total joint replacement, high concentrations of mesenchymal stromal cells (MSCs) are released at the implantation site. They can be found in cell-tissue composites (CTC) that are regularly removed by surgical suction. A surgical vacuum suction handle was filled with bone substitute granules, acting as a filter allowing us to harvest CTC. The purpose of this study was to investigate the osteopromotive potential of CTC trapped in the bone substitute filter material during surgical suction. In the course of 10 elective total hip and knee replacement surgeries, β-tricalcium-phosphate (TCP) and cancellous allograft (Allo) were enriched with CTC by vacuum suction. Mononuclear cells (MNC) were isolated from the CTC and investigated towards cell proliferation and colony forming unit (CFU) formation. Furthermore, MSC surface markers, trilineage differentiation potential and the presence of defined cytokines were examined. Comparable amounts of MNC and CFUs were detected in both CTCs and characterized as MSC‱ of MNC with 9.8 ± 10.7‱ for the TCP and 12.8 ± 10.2‱ for the Allo (p = 0.550). CTCs in both filter materials contain cytokines for stimulation of cell proliferation and differentiation (EGF, PDGF-AA, angiogenin, osteopontin). CTC trapped in synthetic (TCP) and natural (Allo) bone substitute filters during surgical suction in the course of a joint replacement procedure include relevant numbers of MSCs and cytokines qualified for bone regeneration.
... At present, the insu cient amount of autologous bone and limited osteogenic ability of conventional bone substitutes have led to research efforts aimed at identifying bone llings with excellent osteogenic properties [7][8][9]. The stem cell Screen-Enrich-Combine(-biomaterials) Circulating System (SECCS), developed previously by us, can rapidly prepare enriched mesenchymal stem cells (MSCs)/beta tricalcium phosphate (β-TCP) composites, which have shown good bone repair ability in animal and various clinical bone defect models [10,11]. However, the e cacy of repair needs to be further veri ed as an in-frame lling to repair complex anatomical bone defects. ...
Background: Repairing complex anatomical load-bearing bone defects is difficult because it requires restoration of load-bearing function, reconstruction of anatomical shape and repair by regenerated bone. We previously developed a Screen–Enrich–Combine(-biomaterials) Circulating System (SECCS) for rapid intraoperative enrichment of autologous bone marrow mesenchymal stem cells (MSCs) to enhance the osteogenic ability of porous bone substitutes. In this study, we prepared a 3D-printed Ti6A14V macroporous frame matching the defect shape to provide early load-bearing support and evaluated the efficacy of filling the frame with SECCS-processed MSCs/beta tricalcium phosphate (β-TCP) for long-term bone growth.
Method: Mechanical testing of the cylindrical Ti6Al4V frame identified optimal 3D printing parameters. The lateral part of a goat distal femur was used as the defect model, and a matching electron beam melting technology-prepared (EBM) Ti6Al4V frame was fitted. Three frames filled with nothing, pure porous β-TCP or SECCS-processed MSCs/β-TCP were fixed onto the defect site. Repair efficacy was evaluated by X-ray radiography, computed tomography, histology and histomorphology.
Results: In the basic regular hexagon printing unit, the combined side width (w) and inscribed circle diameter (d) determines the printing frame’s mechanical strength. The compressive load was significantly higher for w = 1.9 mm, d = 4.4 mm than for w = 1.7 mm, d = 4.0 mm or w = 2.0 mm, d = 5.0 mm (P < 0.05). The EBM-prepared Ti6Al4V defect-matched frame was well maintained 9 months after implantation. The MSCs successfully adhered to the wall of the porous β-TCP in the SECCS-processed group and had spread fully in the test samples. Each goat in the MSCs/β-TCP–filling group had approximately 31,321.7 ± 22,554.7 MSCs and a larger area of new bone growth inside the frame than the areas in the control and blank groups.
Conclusion: Filling the 3D-printed Ti6Al4V large-aperture frame with osteogenic materials achieved biological reconstruction over a larger area of regenerated bone for repair of complex anatomical weight-bearing bone defects under the condition of early frame-supported load bearing. MSCs/β-TCP prepared by SECCS can be used as a filling material for this type of bone defect to obtain more efficacious bone repair.
... Mesenchymal stem cell enrichment technology focuses on techniques for isolating sufficient amounts of MSCs using nonculturing methods. Presently, the primary methods of MSC enrichment include density gradient centrifugation and the use of a screen-enrich-combine circulating system for MSCs [12][13][14][15][16]. The former method obtains a bone marrow nucleated cell layer rich in MSCs via density gradient centrifugation of the bone marrow [12,17]. ...
... The former method obtains a bone marrow nucleated cell layer rich in MSCs via density gradient centrifugation of the bone marrow [12,17]. By using a porous material as a filtration medium, the latter method, at the time of filtration, utilizes the quick adhesion of MSCs to the inner and outer surfaces of porous materials during filtration because of the rapid adhesion characteristic of MSCs [13,14]. Consequently, the cell-material combination is directly achieved. ...
... Important goals in the field of orthopedic research have been to develop bone repair materials with improved osteogenetic capability, osteoinductivity, and osteoconductivity and to become less dependent on the use of autologous bones [19,20]. Because MSCs play indispensable roles in bone repair, several cell-processing strategies have been used for MSC extraction and their combination with traditional bone repair materials to enhance their osteogenic capacity [4,12,13,[21][22][23]. The application of non-in vitro culture techniques can circumvent some ethical and technical limitations. ...
Background:
When bone marrow is repeatedly filtered through porous material, the mesenchymal stem cells (MSCs) in the bone marrow can adhere to the outer and inner walls of the carrier material to become enriched locally, and this is a promising method for MSC enrichment. In this process, the enrichment efficiency of MSCs involved in the regulation of the cell ecology of postfiltration composites containing other bone marrow components is affected by many factors. This study compared the enrichment efficiency and characterized the phenotypes of enriched MSCs obtained by the filtration of autologous bone marrow through different porous bone substitutes.
Methods:
Human bone marrow was filtered through representative porous materials, and different factors affecting MSC enrichment efficiency were evaluated. The soluble proteins and MSC phenotypes in the bone marrow before and after filtration were also compared.
Results:
The enrichment efficiency of the MSCs found in gelatin sponges was 96.1% ± 3.4%, which was higher than that of MSCs found in allogeneic bone (72.5% ± 7.6%) and porous β-TCP particles (61.4% ± 5.4%). A filtration frequency of 5-6 and a bone marrow/material volume ratio of 2 achieved the best enrichment efficiency for MSCs. A high-throughput antibody microarray indicated that the soluble proteins were mostly filtered out and remained in the flow through fluid, whereas a small number of proteins were abundantly (> 50%) enriched in the biomaterial. In terms of the phenotypic characteristics of the MSCs, including the cell aspect ratio, osteogenetic fate, specific antigens, gene expression profile, cell cycle stage, and apoptosis rate, no significant changes were found before or after filtration.
Conclusion:
When autologous bone marrow is rapidly filtered through porous bone substitutes, the optimal enrichment efficiency of MSCs can be attained by the rational selection of the type of carrier material, the bone marrow/carrier material volume ratio, and the filtration frequency. The enrichment of bone marrow MSCs occurs during filtration, during which the soluble proteins in the bone marrow are also absorbed to a certain extent. This filtration enrichment technique does not affect the phenotype of the MSCs and thus may provide a safe alternative method for MSC enrichment.
... Furthermore, cell-seeded scaffolds were found to be superior to autograft, allograft or cell-seeded allograft in terms of bone formation at ectopic implantation sites 298 . An innovative appliance named the stem cell screen-enrich-combine(-biomaterials) circulating system (SECCS) was designed in another study 299 . In that study, 42 patients who required bone graft underwent SECCS-based treatment. ...
... The results showed 85.53 % ± 7.95 % autologous mesenchymal stem cells were successfully screened, enriched, and seeded on the bioceramic scaffolds synchronously. Clinically, all patients obtained satisfactory bone healing 299 . ...
In the late 1960s, strong interest grew in studying various types of ceramics as potential bone grafts thanks to their suitable biomechanical properties. A bit later, such synthetic biomaterials were termed bioceramics. Since then, there has been a number of important achievements in this field. Namely, after the initial development of bioceramics that were just tolerated in the physiological environment, emphasis was shifted towards those able to form direct chemical bonds with the adjacent bones and tissues. Afterwards, based on selection of the appropriate chemical composition coupled with structural and compositional controls, it became possible to choose whether the bioceramic implants remained biologically stable once incorporated into the skeletal structure or whether they should be resorbed over time. At the turn of the millennium, a new concept of regenerative bioceramics was developed and such formulations became an integrated part of the tissue engineering approach. Now bioceramic scaffolds are designed to induce bone formation and vascularization. These scaffolds are usually porous and often harbor various biomolecules and/or cells. This review describes the major types and properties of bioceramics suitable for tissue engineering.
